7PTJ

C54S mutant of choline-sulfatase from E. meliloti CECT4857 bound to HEPES

Summary for 7PTJ

• Entry DOI: 10.2210/pdb7ptj/pdb
• Related: 6G5Z 6G60
• Descriptor: Choline sulfatase, 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID, SULFATE ION, ... (6 entities in total)
• Functional Keywords: choline, sulfatase, sulfate, hydrolase
• Biological source: Rhizobium meliloti (Ensifer meliloti, Sinorhizobium meliloti)
• Total number of polymer chains: 4
• Total formula weight: 242148.46

Authors

Gavira, J.A.,Martinez-Rodriguez, S. (deposition date: 2021-09-27, release date: 2022-08-03, Last modification date: 2024-01-31)

Primary citation

Gavira, J.A.,Camara-Artigas, A.,Neira, J.L.,Torres de Pinedo, J.M.,Sanchez, P.,Ortega, E.,Martinez-Rodriguez, S.
Structural insights into choline-O-sulfatase reveal the molecular determinants for ligand binding.
Acta Crystallogr D Struct Biol, 78:669-682, 2022

PubMed Abstract: Choline-O-sulfatase (COSe; EC 3.1.6.6) is a member of the alkaline phosphatase (AP) superfamily, and its natural function is to hydrolyze choline-O-sulfate into choline and sulfate. Despite its natural function, the major interest in this enzyme resides in the landmark catalytic/substrate promiscuity of sulfatases, which has led to attention in the biotechnological field due to their potential in protein engineering. In this work, an in-depth structural analysis of wild-type Sinorhizobium (Ensifer) meliloti COSe (SmeCOSe) and its C54S active-site mutant is reported. The binding mode of this AP superfamily member to both products of the reaction (sulfate and choline) and to a substrate-like compound are shown for the first time. The structures further confirm the importance of the C-terminal extension of the enzyme in becoming part of the active site and participating in enzyme activity through dynamic intra-subunit and inter-subunit hydrogen bonds (Asn146-Asp500-Asn498). These residues act as the `gatekeeper' responsible for the open/closed conformations of the enzyme, in addition to assisting in ligand binding through the rearrangement of Leu499 (with a movement of approximately 5 Å). Trp129 and His145 clamp the quaternary ammonium moiety of choline and also connect the catalytic cleft to the C-terminus of an adjacent protomer. The structural information reported here contrasts with the proposed role of conformational dynamics in promoting the enzymatic catalytic proficiency of an enzyme.

PubMed: 35503214
DOI: 10.1107/S2059798322003709

Experimental method

X-RAY DIFFRACTION (2.1 Å)